双控激活式细胞核定位双光子荧光探针研究

It is one of the core content of researches in analytical chemistry that development of high efficient and accurate technique for acquiring chemical and biological information. Focusing on the forefront of the international science arena in bio-analysis and chemo/biosensing, this project would carry out original research on the basis of exploring subcelluar informations with new principle and new method: screening and synthesis of nuclear targetable fluorescent probe for important bio-relevant species such as metal ions, reactive oxygen species, reactive nitrogen species, reactive sulfur species, pH, et al; Inspiring from the characteristic component of cellular nucleus-DNA, design of targetable group and recognition site based on the special interactions between DNA and organic small molecule and selective chemical reactions; construction of the new system of nuclear location fluorescent analysis by combing the strategy of the simultaneous DNA and analyte activatable signal output with two-photon fluorescent technique. The execution of this project would provide new idea to develop and enrich the types of subcellular location fluorescent probe, and give new tool and new method to explore the relation between the biological active species with life activities and diseases in subcellular level.

发展准确高效的生化信息获取技术，是当前分析化学研究的核心内容之一。本项目以亚细胞层面生化信息获取新原理与新方法为研究基点，瞄准生命分析和化学生物传感国际前沿领域，开展原始创新的基础研究：针对细胞核内重要生命活性物质（如金属离子，活性氧物种，活性氮物种，活性硫物种，PH值等），筛选合成高特异性和强亲和力的靶向细胞核荧光分子探针工具；从细胞核内特有组分脱氧核糖核酸(DNA)寻找突破点，基于DNA/小分子特异性相互作用和选择性化学反应，设计靶向基团和专一性分子识别单元; 运用DNA与分析对象双重控制激活式信号输出策略，结合双光子荧光技术优势，系统构建细胞核区域定位荧光可视化分析新体系。本项目的执行将为丰富与发展高效亚细胞区域定位荧光探针提供新思路，为探究亚细胞水平上生物活性物质变化与生命活动规律或疾病成因之间关系提供新工具和新方法。

本项目以发展高效实时原位生化信息获取新原理与新技术为出发点，针对生命分析化学关键科学问题，围绕新型双光子荧光传感成像分析体系建立，系统研究了脱氧核糖核酸(DNA)-染料相互作用机制，筛选出具有DNA嵌入荧光增强特性的双光子荧光团，并发展了一类目标物诱导DNA/染料结合增强型双光子荧光生化分析新方法；基于双控激活式策略，通过协同调控双光子荧光共振能量转移(TP-FRET) 和分子内价电荷转移(ICT) 信号传导机制，构建了适用于复杂体系和亚细胞区域定位成像应用的高效双光子荧光比率传感成像平台；基于选择性化学反应策略，发展了专一通用型分子识别新单元；针对活体、组织、或细胞器内过氧化氢、半胱氨酸、次硝酸、多硫化氢、二氧化硫衍生物等重要生命活性物质和疾病关键标志物，设计合成了系列双光子荧光成像探针。项目执行丰富拓展了荧光可视化分析体系与方法学，为细胞信号通路研究和生物医学成像分析提供了新工具。